CAJ | 학술논문

目的：探讨硫化氢(H2S)是否通过改变葡萄糖调节蛋白78(GRP78)的表达参与其对抗6-羟基多巴胺(6-OHDA)诱导PC12细胞损伤的保护作用。方法应用具有神经毒性的6-OHDA损伤PC12细胞为帕金森病细胞模型，以硫氢化钠(NaHS)作为H2S的供体；应用CCK-8比色法检测细胞存活率；DFCH-DA染色检测细胞内活性氧(ROS)的水平；Rh123染色检测细胞线粒体膜电位(MMP)；Western blot检测GRP78的表达。结果200μmol/L的6-OHDA引起PC12细胞的存活率显著降低，ROS生成增加及MMP降低，且诱导了GRP78的高表达。应用25~400μmol/L的NaHS预处理30 min，呈浓度依赖性抑制6-OHDA引起的细胞存活率降低，其中400μmol/L的NaHS作用最明显，此浓度也可以显著减少6-OHDA引起的ROS增多，提高MMP，同时明显抑制6-OHDA诱导的GRP78高表达。结论 H2S具有抗6-OHDA氧化应激损伤的PC12细胞保护作用，抑制内质网应激分子GRP78的表达可能是其机制之一。
목적：탐토류화경(H2S)시부통과개변포도당조절단백78(GRP78)적표체삼여기대항6-간기다파알(6-OHDA)유도PC12세포손상적보호작용。방법응용구유신경독성적6-OHDA손상PC12세포위파금삼병세포모형，이류경화납(NaHS)작위H2S적공체；응용CCK-8비색법검측세포존활솔；DFCH-DA염색검측세포내활성양(ROS)적수평；Rh123염색검측세포선립체막전위(MMP)；Western blot검측GRP78적표체。결과200μmol/L적6-OHDA인기PC12세포적존활솔현저강저，ROS생성증가급MMP강저，차유도료GRP78적고표체。응용25~400μmol/L적NaHS예처리30 min，정농도의뢰성억제6-OHDA인기적세포존활솔강저，기중400μmol/L적NaHS작용최명현，차농도야가이현저감소6-OHDA인기적ROS증다，제고MMP，동시명현억제6-OHDA유도적GRP78고표체。결론 H2S구유항6-OHDA양화응격손상적PC12세포보호작용，억제내질망응격분자GRP78적표체가능시기궤제지일。
Objective To investigate whether hydrogen sulfide (H2S)involved in the protection of PC12 cells against 6-hydroxydopamine(6-OHDA)-induced injury by changing the glucose-regulated protein 78(GRP78)expression. Meth-ods 6-OHDA was used to establish the Parkinson disease model in PC12 cells with dopaminergic neurons characteris-tics.Sodium hydrosulfide(NaHS)was used as a H2S donor.The viability of PC12 cells was measured by CCK-8 assay.The level of reactive oxygen species (ROS)in PC12 cells was measured by DCFH-DA staining.The mitochondrial membrane potential (MMP)was analyzed by rhodamine 123 staining.The expression of GRP78 was evaluated by Western blot. Re-sults 200 μmol/L 6-OHDA induced a decrease in cell viability and overproduction of ROS as well as dissipation of MMP in PC12 cells.6-OHDA induced the upregulation of GRP78 expression.When PC12 cells were treated with NaHS 30 min before 6-OHDA treatment a decrease in viability of PC12 cells induced by 6-OHDA was concentration-depen-dently blocked by NaHS (25-400μmol/L).Pretreatment with NaHS at 400μmol/L obviously inhibited the dissipation of MMP and overproduction of ROS induced by 200 μmol/L 6-OHDA.Furthermore,NaHS preconditioning obviously di-creased the upregulation of GRP78 expression induced by 6-OHDA. Conclusion H2S protected PC12 cells against 6-OHDA-induced oxidative stress injury and inhibiting the expression of GRP78 may be one of the mechanism under-lying cytoprotection induced by H2S preconditioning.